LOCUS       sequence-398287-        7177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express WT-LAT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7177)
  AUTHORS   Peruzzi JA, Gunnels TF, Edelstein HI, Lu P, Baker D, Leonard JN, 
            Kamat NP
  TITLE     Enhancing extracellular vesicle cargo loading and functional 
            delivery by engineering protein-lipid interactions.
  JOURNAL   Nat Commun. 2024 Jul 4;15(1):5618. doi: 10.1038/s41467-024-49678-z.
  PUBMED    38965227
REFERENCE   2  (bases 1 to 7177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2024-07-4"; volume: "15(1):5618. doi"; pages: " 
            10.1038/s41467-024-49678-z"
COMMENT     Sequence Label: pJPT018-LAT_WT-HaloTag-3x FLAG
FEATURES             Location/Qualifiers
     source          1..7177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             148..1032
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1042..1107
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1160..1281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1171..1190)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1225..1244
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1396..1442
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1472..1608
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1653..1724
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1950..2538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2030..2049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3038..4247
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4084..4103
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4251..4850
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4251..4270)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4747..4767
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4877..4925
                     /note="synthetic polyadenylation signal"
     CDS             join(5191..5259,5260..6051)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5409..5428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6064..6200
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6352..6423
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6428..6474)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6492..6540
                     /note="synthetic polyadenylation signal"
     enhancer        6621..6924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6926..7128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7079..7099
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg gaggaggcca tcctggtccc ctgcgtgctg
       61 gggctcctgc tgctgcccat cctggccatg ttgatggcac tgtgtgtgca ctgccacaga
      121 ctgccaggct ccggatccgg cggctccgaa atcggtactg gctttccatt cgacccccat
      181 tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc
      241 cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg
      301 catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac
      361 aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa
      421 gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc
      481 cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc
      541 cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc
      601 accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg
      661 ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc
      721 ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt
      781 gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct
      841 gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct
      901 cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg
      961 ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg
     1021 gagatttccg gctccgaatt cgattacaag gaccacgatg gcgactataa ggatcacgac
     1081 atcgactaca aggacgatga cgacaagtga gctagcttga ctgactgaga tacagcgtac
     1141 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1201 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1261 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1321 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1381 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1441 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1501 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     1561 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     1621 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     1681 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     1741 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     1801 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     1861 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     1921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2761 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     2821 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     2881 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     2941 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3001 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     3061 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     3121 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     3181 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     3241 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3301 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3361 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     3421 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     3481 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     3541 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     3601 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     3661 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     3721 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     3781 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     3841 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     3901 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     3961 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     4021 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     4081 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     4141 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     4201 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     4261 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4321 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4381 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4441 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4501 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4561 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4621 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4681 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4741 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4801 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     4861 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     4921 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     4981 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5041 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5101 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5161 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5221 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5281 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5341 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5401 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5461 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5521 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5581 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5641 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5701 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     5761 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     5821 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5881 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5941 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6001 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6061 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6121 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6181 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6241 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6301 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6361 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6421 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6481 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     6541 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     6601 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     6661 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     6721 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     6781 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     6841 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     6901 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
     6961 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     7021 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     7081 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     7141 cgtcagatca gatctttgtc gatcctacca tccactc
//
